A 1H-1,5-benzodiazepine derivative induces apoptosis in colorectal cancer cells by inhibiting JAK/STAT signaling pathway

Colorectal cancer (CRC) is among the most common cancers worldwide, with limited pharmacological options currently. Benzodiazepine and its synthetic derivatives are now of interest as their bioactive attributes, especially their inhibitory effect on various cancer cell lines. Herein we designed and synthesized of a series of compounds, among which T0, a 1H-1,5-benzodiazepine derivative, which exhibited potential inhibitory activity against colorectal cell lines in vitro. Relative to 5-fluorouracil, compound T0 exhibited a superior cytotoxic effect while maintaining lower toxicity. Additionally, our research indicated that T0 decreased mitochondrial membrane potential and activated reactive oxygen species (ROS) production in CRC cells leading to mitochondria-mediated apoptosis. Mechanistically, T0 could downregulate the JAK/STAT signaling pathway which was in good agreement with docking result and the observed in vitro activity. Structural analysis through molecular docking and cellular thermal shift assay (CESTA) revealed T0's high-affinity interaction with the protein JAK, suggesting T0 was a potential JAK inhibitor. Overall, this study suggested T0 may be a potential lead compound for the therapeutic option of human CRC.